Development of Aflatoxin B1-Lysine Adduct Monoclonal Antibody for Human Exposure Studies

Jia Sheng Wang, Salahaddin Abubaker, Xia He, Guiju Sun, Paul Timothy Strickland, John Davis Groopman

Research output: Contribution to journalArticle

Abstract

Mouse monoclonal antibodies were developed against a synthetic aflatoxin B1 (AFB)-lysine-cationized bovine serum albumin conjugate. The isotype of one of these antibodies, IIA4B3, has been classified as immunnglobulin G1(λ). The affinity and specificity of IIA4B3 were further characterized by a competitive radioimmunoassay. The affinities of IIA4B3 for AFB and its associated adducts and metabolites are ranked as follows: AFB-lysine > 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB > AFB = 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB > aflatoxin M1 > aflatoxin Q1. IIA4B3 had about a 10-fold higher affinity for binding to AFB-lysine adduct than to AFB when 3H-AFB-lysine was used as the tracer. The concentration for 50% inhibition for AFB-lysine was 0.610 pmol; that for AFB was 6.85 pmol. IIA4B3 had affinities at least sevenfold and twofold higher than those of 2B11, a previously developed antibody against parent AFB, for the major aflatoxin-DNA adducts 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB and 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB, respectively. An analytical method based on a competitive radioimmunoassay with IIA4B3 and 3H-AFB-lysine was validated with a limit of detection of 10 fmol of AFB-lysine adduct. The method has been applied to the measurement of AFB-albumin adduct levels in human serum samples collected from the residents of areas at high risk for liver cancer.

Original languageEnglish (US)
Pages (from-to)2712-2717
Number of pages6
JournalApplied and Environmental Microbiology
Volume67
Issue number6
DOIs
StatePublished - Jun 2001

Fingerprint

Aflatoxin B1
aflatoxin B1
antibody
monoclonal antibodies
lysine
Monoclonal Antibodies
serum
Lysine
cancer
analytical method
metabolite
tracer
fold
DNA
exposure
aflatoxin B1-lysine adduct
Radioimmunoassay
radioimmunoassays
aflatoxins
Aflatoxin M1

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science(all)
  • Microbiology

Cite this

Development of Aflatoxin B1-Lysine Adduct Monoclonal Antibody for Human Exposure Studies. / Wang, Jia Sheng; Abubaker, Salahaddin; He, Xia; Sun, Guiju; Strickland, Paul Timothy; Groopman, John Davis.

In: Applied and Environmental Microbiology, Vol. 67, No. 6, 06.2001, p. 2712-2717.

Research output: Contribution to journalArticle

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abstract = "Mouse monoclonal antibodies were developed against a synthetic aflatoxin B1 (AFB)-lysine-cationized bovine serum albumin conjugate. The isotype of one of these antibodies, IIA4B3, has been classified as immunnglobulin G1(λ). The affinity and specificity of IIA4B3 were further characterized by a competitive radioimmunoassay. The affinities of IIA4B3 for AFB and its associated adducts and metabolites are ranked as follows: AFB-lysine > 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB > AFB = 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB > aflatoxin M1 > aflatoxin Q1. IIA4B3 had about a 10-fold higher affinity for binding to AFB-lysine adduct than to AFB when 3H-AFB-lysine was used as the tracer. The concentration for 50{\%} inhibition for AFB-lysine was 0.610 pmol; that for AFB was 6.85 pmol. IIA4B3 had affinities at least sevenfold and twofold higher than those of 2B11, a previously developed antibody against parent AFB, for the major aflatoxin-DNA adducts 8,9-dihydro-8-(N7-guanyl)-9-hydroxy-AFB and 8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl formamido)-9-hydroxy-AFB, respectively. An analytical method based on a competitive radioimmunoassay with IIA4B3 and 3H-AFB-lysine was validated with a limit of detection of 10 fmol of AFB-lysine adduct. The method has been applied to the measurement of AFB-albumin adduct levels in human serum samples collected from the residents of areas at high risk for liver cancer.",
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